http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
- 中文 을 입력하시려면 zhongwen을 입력하시고 space를누르시면됩니다.
- 北京 을 입력하시려면 beijing을 입력하시고 space를 누르시면 됩니다.
RISS 인기검색어
- 검색키워드
- 주제어 : 원형질체 융합 (검색결과 78 건)
-
Penicillium verruculosum의 分生子 原形質體融合
섬유소 분해효소 생산균 Penicilium verruculosum F-3 의 균주개량을 목적으로 분생자원형질체의 생성, 재생, 융합 및 융합체의 유전적 안전성, Cellulase 생산성 등을 검토 하였다. 2 -DG (25 ug/㎖l)를 첨가한 최소액체 배지에서 10∼12 시간 전배양한 conidia를 1% Novozyme 234로 30℃, 3시간 처리시에 원형질체 생성율이 가장 높았다. 원형질체의 생성 및 재생을 위한 삼투압 안정제로는 각각 0.6M (NH₄) ₂SO₄및 0.6M MgSO₄가 효과적 이었다. 영양요구성 유래의 원형질체의 생성율, 재생율 및 융합율은 각각 48∼51%, 34.5~49.2%및 2.2 ×10^-2 ∼ 3.4×10^-l 수준 이었다. 융합체의 자연분리, DNA 함량측정, 핵염색 등의 비교 검토를 통해 핵융합이 이루어졌음을 확인하였고 이들의 핵형은 이수체이며 5개월간의 계대배양 후에도 유전적 안전성이 매우 높았다. Cellulase 고생산융합체로 선별된 4주의 Cellulase활성은 융합모균이나 야생주에 비해 1.3 ∼ 4배나 높았다. 이들 융합체중 Cellulase 활성이 가장 높은 FPV 1015A3은 본 효소 생성이 최고치에 달하는 시간이 융합모균이나 야생주에 비해 1/2 ∼ 1/3로 단축됨을 보여 차후 본효소 유도기구 해명을 위한 분자적 차원에서의 연구에 좋은 재료가 될수 있음이 인정되었다. Optimal conditions for the intraspecific protoplast fusion using conidial protoplasts of Penicillium verruculosum F-3, one of the hypercellulolytic enzyme-producing fungi, were studied in order to investigate the possibility of strain improvement for cellulolytic enzyme productivity via intraspecific protoplast fusion. Among the various commercial cell wall lytic enzymes tested, l%(w/v) Novozyme234, when the reaction was performed at 30˚C for 3hrs, was the most effective for protoplast isolation. High protoplast yield were obtained from the preincubated swollen conidiospores cultured in the liquid minimal medium containing 2-deoxy-D-glucose(25ug/ml) for 10 - 12hrs. The best osmotic stabilizers for the isolation and regeneration of protoplasts were 0.6M (NH₄)₂SO₄ and 0.6M MgSO₄, respectively. The protoplast yield of parent strains for fusion ranged from 48% to 51%, regeneration frequency of the protoplasts from the conidiospores in RCM ranged from 34.5 to 49.2%, and reversion frequency in RMM ranged from 1.5 × 10^-9 to 7.8 × 10^-7. The intraspecific fusion frequency between protoplasts from auxotrophs ranged from 2.2 × 10^-2 to 3.4 × 10^-1 in 30% PEG-10mM CaCl₂-0.05M citrate buffer(pH 5.6) system. From the observation of spontaneous segregation, measurement of DNA content, and nuclear staining of the cellulolytic fusants with Giemsa, it was confirmed that nuclear fusion occured in the fusants, their karyotypes were aneuploid, and their genetic stability was very high at least for 5 months' subculture. The four non-parental prototrophic hybrids selected for cellulase productivity revealed cellulolyitc activity about 1.3 - 4 folds of the wild or parental strains. Especially one hybrid FPV1015A3, which had the highest cellulolytic activity among them required induction period only one thirds or half of that required by wild or parental strains. The strain, therfore, was expected to be used as a good material for study of the cellulase induction mechanism at molecular level.
-
酵母, Phaffia rhodozyma의 原形質體 融合
Astaxanthin 을 생산하는 효모, Phaffia rhodozyma 로부터 제조된 상보적 돌연번이 균주사이의 세포 융합을 통하여 astaxanthin 을 대량 생산하는 균주를 얻고자 시도 하였다. 돋언번이주들의 원형질체 형성수율은 19 - 81 % 이었으며 이들의 원형질체 융합율은 1.3 × 10^(-5)에서 6.0 × 10^(-5) 이었고 DNA 함량, 핵 염색, UV 조사에 대한 생존력 비교 그고고 형질분리 분석 결과 재 조합형의 분리로 인하여 핵융합을 확인 하였다. 이 융합체들은 비 선택배지에서 1 년간 보존 후에도 유전적으로 안정하였으며, 이들 중 F 16 은 야생형의 모 균주와 비교했을때 astaxanthin 함유량이 약 7배 증가하였다. To obtain astaxanthin-overproducing strains by protoplast fusion technique, the cell fusion between complementary mutants isolated from the astaxanthin--producing yeast Phaffia rhodozyma was carried out. The rate of protoplast formation ranged from 19 to 81 %. Protoplast fusion showed with a frequency range of 1.3 X 10^(-5) to 6.0 x 10^(-5), and nuclear fusion in the cells of hybrids was demonstrated by several techniques such as the isolation of recombinants after mitotic segregation of parental genetic markers, the estimation of DNA content, the direct observation of nuclei with nuclear staining, and the comparison of survival rate to UV exposure. Several hybrids tested maintained their ploidy and ataxanthin - producing activity after keeping for 12 months even under nonselective conditions. One those hybrids, fusant F16, showed approximately 7-fold increase in astaxanthin content when compared with wild parent.
-
-
-
Brevibacterium lactofermentum의 원형질체 융합에 의한 유전자 재조합
이혜경 淑明女子大學校 大學院 1989 국내석사
Genetic manipulation by protoplast fusion B. lactofermentum ATCC 13869 was applied for genetic mapping on the chromosome. As a genetic selection marker, the auxotrophic mutants, B. lactofermentum SWA (arg^- trp^-) and B. lactofermentum SWB (met^- ser^-) were obtained from UV and NTG treatment. The rate of protoplast formation was 99.93% ∼ 99.98% when strains were treated with 0.3 unit/ml of penicillin G in mid exponential growth phase for 1.5 hours, followed incubation wi th 400㎍/ml of lysozyme in lysis fluid supplemented with 0.4M sucrose as osmotic stabilizer (pH 7). Frequency of protoplast regeneration in B. lactofermentum SWA and B. lactofermentum SWB was 9.27% and 10.32% respectively, on regeneration medium containing 0.5M sodium succinate, 50mM Mg^2+, and 3% PVP. Intact or fused protoplasts were placed by placing the suspension on the surface of the regeneration agar medium, and 4ml of the same medium with 0.4% low melting point agarose at 38℃ was poured onto agar plate. In intraspecific protoplast fusion between B. lactofermentum SWA and B. lactofermentum SWB, fusion frequency of 2.30 × 10^-5 per regenerated cells was observed by using the 100mM CaCl₂ and PEG 6,000, 30%(W/V) in fusion fluid. Relative recombinant frequencies in each marker by means of selective media could be used for genetic analysis and gene mapping.
-
原形質體融合體 選拔을 위한 β-Glucuronidase 標識의 당근에로의 導入 및 發現
To select somatic hybrids of carrot and ginseng by protoplast fusion, E. coli β-glucuronidase (GUS) gene as a marker was introduced to carrot genome by using a binary vector system of Agrobacterium tumefaciens. Protoplasts were enzymatically isolated from carrot suspension culture and regenerated into whole plants via somatic embryogenesis after 8 weeks of culture. Protoplasts of carrot and ginseng labeled with fluorcsccin isothiocyanate and rhodamine isothiocyanate, respectively, were fused by polyethylene glycol. An intermediate stage in fusion was observed under cpiscopic illumination. pBI 121 harboring CaMV 35s promoter-GUS fusion was introduced into A. tumefaciens to transform carrot. Hypocotyl and cotyledon explants were co-cultivated with A. tumefaciens carrying pBI 121 on a medium containg 200 ㎎/ℓ kanamycin. Kanamycin-resistant calli were formed after 3 to 4 weeks of culture. Production of agropine and mannopinc was detected in the resistant calli by paper electrophoresis. Southern and Northern blotting analyses confirmed that the resistant calli were transformed with GUS gene. High GUS activities were detected in transformed calli by spectrophotometric and histochemical assays. Roots were sporadically induced from a few of the calli, and a preliminary histochemical assay revealed that GUS gene under CaMV 35s promoter was preferentially expressed in the vascular bundle and the lateral root primordium compared to the hair and the apical meristem.
-
-
-
-
연관 검색어 추천
이 검색어로 많이 본 자료
활용도 높은 자료
